The effects of restraint on the amount of nondialysable radioactive sulfate in the gastric wall and the gastric juice and saliva were investigated. It was found that restraint provokes a significant decrease in salivary radioactive sulfate. This, in turn, is responsible for the decrease of sulfate in the gastric contents observed under these conditions in rats with pyloric ligation. Esophageal ligation associated with this prevents passage of saliva and lowers the amount of radioactive sulfate in the gastric juice. Restraint causes then an increase in the amount of sulfate in the gastric juice, the value observed being very much lower than that of rats with a free esophagus. At the level of the gastric wall, the change observed during restraint does not reach a significant threshold

Chayvialle, J. A.

Lambert, R.

Ruet, D.

English

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THE EFFECTS OF RESTRAINT ON UPTAKE OF RADIOACTIVE SULFATE
IN THE SALIVARY AND GASTRIC SBCRETIONS OF RATS WITH PYLORIC LIGATION
J. A. Chayvialle, R. Lambert, D. Ruet
Translation of "Effets de la constrainte sur V incorporation du radiosulfate
dans les secretions salivaires etgastriques:du rat a pylore lie", -Comptes
Rendus ses Seances de la Societe de Biologie et de sea Filiales, Vol. 166,
No. 1, 1919, pp 114-118 	
I
80
NATIONAL AERONAUTICS AND SPACE ADMINISTRATION
WASHINGTON, D. C. 20546	 MARCH 1980
EFFECTS OF RESTRAINT ON THE INCORPORATION OF RADIOACTIVE SULFATE
IN THE SALIVARY AND GASTRIC SECRETIONS OF RATS WITH PYLORIC LIGATION
J. A. Chayvialle, R. Lambert and D. Ruet
	 /114*
_
	
	 Restraint provokes in the rat a diminution of sulfated mucous
substances present in the glandular portion cf the gastric wall.
A relationship between this variation and ulcerogenesis has been
suggested [1,2]. Radioactive sulfate is incorporated specifically
in the form of eater-sulfates in these mucous substances. The
reduction of this incorporation in the restrained animal can be due
either to a decrease of the synthesis of the ester-sulfates in
glandular mocosa, or to an increase in their excretion into the
gastric juice. Simultaneous study of t:ie radioactive sulfate in the
wall and in the contents of the stomach should make it possible to
decide between these two hypotheses. It is necessary, however, not
to exclude contamination of the gastric juice by saliva. This work
is dedicated to the study of the effects of restraint on the amount
of non-dialysable radioactive sulfate in the gastric wall, in the
gastric juice and in saliva. Various procedures have been employed
`	 in order to collect these two secretions separately.
Method. 102 Wistar rats (86 males, 180-200gm, and 16 females,
200-250 gm) received at the beginning of the experiment 100 VC of
sulf#"r 35 by the i.p. route (sodium sulfate, C.E.A.). They were
divided i-to three groups accordii.g to the surgical protocol which
always included pyloric ligation by the ihay technique 131: Group I:
gastric secretion was collected in 60 rats, either after isolated 	 /115
pyloric ligation (group l.a: 29 animals), or after pyloric and eso-
Numbers in the margin indicate pagination in original foreign text.
1
fl .. ,
s
phageal ligation (group l.b: 31 animals), thus eliminating contam-
ination from saliva. The contents of the stomach and the gastric
glandular wall were analyzed simultaneously. Group II: in 16 rats
a ligature was placed the length of the border between the rumen and
the glandular stomac'. The salivary secretion thus was collected in
a pure state in the rumen. Group III: in 26 rats, the saliva was
collected directly by means of a catheter maintained in the abdominal
esophagus by an esophageal ligature and connected to an intraperit-
oneal small balloon. In each group, the animals were strictly paired
according to weight and sex: a control in the cage, the other rat
restrained in a metallic tube [4].
The rats were sacrificed with ether at the 6th hour. The stomach
or the rumen Here opened and washed with 20 ml of 0.3M sodium chloride
(pH = 1.5). 'The gastric glandular wall was separated from the rumen
and ground up. About 1110 of the tissue was sampled for determina-
tion of relation of the dry weight to the fresh weight (weighed
before and after dessication at 100°C for 24 hours). The remainder
of the glandular wall was placed in 20 ml 0.3M sodium chloride
(pH = 1.5) and subjected to the same analysis time as the secretions:
proteolysis by pepsin (Worthington twice-crystallized pepsin, 20 mg
per sample, 24 hours at 37 0C), dialysis in running water for 48 hours
after addition of 2 ml 5% sodium sulfate for comparison (laboratory
temperature). The radioactivity was determined at the end of the
dialysis by a liquid scintillation counter (Packard Tricarb 3375).
After correction for quenching and loss, the results were expressed
in dpm. The standard deviation of the average was calculated for
each value and indicated in the tables. Non-dialysable radioactive
sulfate after peptic proteolysis is considered the index of incorpor-
ation of the isotope in the sulfated glycoproteins and polysaccharides.
The different series were compared by the Student t test, the
threshold of probability being fixed at 5%.
Results. - 1. When the gastric contents are collected after
isolated pyloric ligation (group I.a.), the restraint provokes a
significant decrease (p < 0.05) of volume and of the quantity of
2
radioactive sulfate in the gastric contents (18.4% of the value for
the controls). Incorporation of the isotope in the wall decreases
but the change is not significant (total amount: 82.5% of the con-
trol value).
2. When an esophageal ligation was performed in the rats with
pyloric ligation (group I.b), the quantity of radioactive sulfate
in the gastric contents decreases in a significant manner (total
amount: 3.7% of the control value, p < 0.01) as well as its concen-
tration per ml (p < 0.01). Restraint provokes a significant increase
(p < 0.05) of the total amount and of the concentration of non-
dialysable radioactive sulfate in the pure gastric juice. The decrease
observed in the wall (total amount: 87.5% of the control value) is
not significant.
3. In rats of group II, bearing a ligature between the rumen 	 /116
and the glandular stomach, restraint causes a significant decrease of
radioactive sulfate in the saliva (total amount: 12.4% of the control
value, p < 0.005. Concentration per ml: 14.2% of the control flue,
p < 0.02) (Table II).
4. When saliva is collected in an isolated manner by esophageal
catheter and balloon (group III), restraint provokes a significant /117
decrease of the salivary radioactive sulfate (total amount: 28.1% of
t
	
	
the control value, p < 0.01. Concentration: 39.3% of the control
value, p < 0.01) (Table II).
Discussion. -
pyloric ligation a
radioactive sulfat,
tents", which then
essentially linked
(total amount: 33%
Restraint for 6 hours provokes in
marked change in the amount of no
e in the combination "gastric wall
reaches 54% of the control value.
to the decrease of sulfate in the
of the control value).
rats with
n-dialysable
+ gastric con-
This change is
gastric contents
The degree of contamination of the gastric juice by salivary
radioactive sulfate is demonstrated by the drop-off of the radio-
3
Controls
	 Restraint
Experimental series Pyl fig/Cerv.
'
Pyl lig Cery
Pyl lig. Esoph. fig.
	 pyl lig. Esoph. lig.
16	 16 13 15No. of rats
Dry weight (mg) 268.7
	 304.9 254.9 263.9
Gastric	 Total amount 349 000	 j 344 000 288 000
i
'301 000
wall	 (dpm 6 hr) (24 200)	 ! (27 300) (29 400) (18 400)
Concentration 1 263	 1 263 1 193 11 099
(d m/m`) 96.5	 130.5 124.7 70.4
Volume (ml) 7.03 1.58 i	 3.18	 ` 2.08
Total amount 615 000 23 000 209 000 59 000
Gastric	 (dpm 6 hr) (176 500) (3 300) (27 5tk) (8 800)
contents
	 Concentration 78 300 19 100 71 200 34 500
(d m/ml) (15 300) (2 400) (10 300) (5 600)
Total amount 911 000 362 600 497 400 360 500
Total	 (d pm 6 hr) (184 300) (28 500)	 1 (46 800) (25 800)
FABLE I.
	 EFFL-CT OF RESTRAINT ON RADIOAC TIVE SULFATE IN THE STOMACH
CO NT 	 AND WALL OF RATS WITH PYLORIC LIGATION, WHETHER SUBJECTED
TO AN ESOPHAGEAL LIGATION OR NOT. IN RATS WITH PYLORIC LIGATION,
RESTRAINT CAUSES A DECREASE OF RADIOACTIVE SULFATE IN THE GASTRIC
JUICE. AFTER ESOPHAGEAL LIGATION, INTRAGASTRIC RADIOACTIVE STILFATE
FALLS AND THE EFFECT OF RESTRAINT TS REVERSED.
4
-_
Groups of experiments Group II	 Group III
Medio astric Ligature Eso ha eal Catheter
Experimental Controls Restraint Controls Restraint
conditions 8 rats 8 rats 15 rats 11 rats
- 3.1 1.8 2.2 1.7
Volume (ml) (1.1)	 l (0.4) (0.2) (0.3)
Dpm16 hr 542 200 67 300 1 190 000 334 200
35 S (191 700) (15 000) 173 800) (107 000)
Dpm/ml 216 600 30 800 557 800 219 000
(71 900) (7 300) (74 600) (63 600)
TABLE II.	 EFFECT OF RESTRAINT ON THE RADIOACTIVE SULFATE IN THE
SALIVA COLLECTED PROXIMAL TO A M,EDIOGASTRIC LIGATURE OR BY AN ESO-
PHAGEAL CATHETER. WHATEVER PROTJCOL IS UTILIZED TO COLLECT THE SALIVA,
RESTRAINT PROVOKES A SIGNIFICANT DECREASE OF THE TOTAL AMOUNT AND OF
THE SALIVARY CONCENTRATION OF RADIOACTIVE SULFATE WITHOUT ANY SIGNI-
FICANT CHANGE IN VOLUME.
activity of the gastric contents af#er esophageal ligation Group
I.b). This is in accord with the decrease of hexosamines in the gas-
tric contents observed after esophageal ligation [5]. The salivary
secretion collected in the pure state (groups II and III) contains a
considerable quantity of radioactive sulfate, the values being more
elevated in group Illbecause of stimulation of salivary secretion by
the intraesophageal catheter. Restraint provokes a marked decrease in
the radioactive sulfate excreted in the saliva (groups II and III).
This change ts in part linked to the immobilization of the animals,
incapable of licking themselves. However, a decrease associated with
synthesis of sulfated glycoproteins in the saliva can be suggested
because the change in the concentration of the radioactive sulfate is
more marked than that in volume.
Pure gastric secretion is collected in rats with esophageal
ligation (group I.b). The quantity of radioactive sulfate present in
the gastric juice is very small. It is necessary, however, t,) take
5
into account a possible inhibitory effect of the esophageal liga-
ture on the gastric glandular mucosa [6]. The size of the change
provoked by restraint in these conditions is modest, but favors an
increased excretion of glycoproteins.
In rats with pyloric ligation, the amount of non-dialysable
radioactive sulfate in the gastric wall falls during restraint (total
amount: 82% of the control value), this change is not significant.
A greater change has previously been reported [1,2] but the experi-
mental protocol was different: analysis was carried out electively
on the radioactive sulfate in the glandular mucosa separated from
the gastric muscular tunic, while our study was carried out on the
entirety of the glandular stomach wall.
In conclusion, restraint provokes a marked decrease in non-
dialysable radioactive sulfate incorporated in the saliva. This
effect is the principal factor in the change in gastric contents of
rats with pyloric ligation subjected to restraint. When the salivary
factor no longer plays a role, an increase of excretion of radio-
active sulfate into the gastric vent is observed under restraint with
no decrease in the sum of "parietal radioactive sulfate + radioactive
sulfate in the secretion".
Summary.	 Study of non-dialysable radioactive sulfate in the
gastric wall and contents in rats should take into account gastric
contamination by salivary sulfated glycoproteins. During six hour
experiments, restraint provokes a significant decrease in salivary
radioactive sulfate. This, in turn, is responsible for the decrease
of sulfate in the gastric contents observed under these conditions in
rat.: with pyloric ligation. Esophageal ligation associated with this
prevents passage of saliva and lowers the amount of radioactive sul-
fate in the gastric juice. Restraint causes then an increase in the
amount of sulfate in the gastric Juice,.the value observed being very
much lower than that of rats with a free esophagus. At the level of
the gastric wall, the change observed during restraint does not reach
a significant threshold within the framework of this study.
6
REFERENCES
[1] C. Andre, M. S. Martin, R. Lambert and F. Martin, C. R. Soo.
Biol., 1959, Vol. 163, P. 162.
[2] R. Lambert, C. Andre and F. Martin, Gastroenterology, 1969,
Vol. 56, p. 200.
[3] H. Shay, S. A. Komarov, S. S. Fels, D. Meranee, M. Cruenstein
and H. Siplet, Gastroenterology, 1945, Vol. 1, p. 43.
[4] R. Lambert, M. S. Martin and F. Martin, C. R. .boo. Biol.,
1957, Vol. 161, p. 816.
[5] K. Kowalewski et al, Amer. J. Dig. Dis., 1909, Vol. 14, P. 788.
[6] D. Brodie and P. Knapp, Gastroenterology, 1966, Vol. 50, P. 787.
If


The effects of restraint on uptake of radioactive sulfate in the salivary and gastric secretions of rats with pyloric ligation

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